Publication Details (including relevant citation information):
Authors: Vishal Gaurav, Susan C. Roberts
Publication date: 2009
Conference name: The American Chemical Society (ACS) Proceedings
Publisher: The ACS National Meeting, Salt Lake City, UT
Paclitaxel, the best-selling anti-cancer drug is a complex diterpenoid synthesized in Taxus plant cell cultures through a multi-step biosynthetic pathway. To unravel the inherent culture heterogeneity and production variability, we are investigating regulation of cellular metabolism through flow cytometric cell sorting technologies. A novel antibody-based indirect immunofluorescent assay was developed to detect paclitaxel accumulation in live plant cells. Concomitantly, fluorescein diacetate assay was performed to ensure cell viability. Experiments were performed with variations in antibody concentrations, wash steps and incubation times to establish an optimal procedure. Subsequently, a novel flow cytometry-based method to quantify the alkaloid content, by investigating the antigenantibody interactions, was developed.